Apparatus for use in luminescence applications

ABSTRACT

Photoluminescence from a sample detector is detected using an array of photo-sensitive detectors. At least one first photo-sensitive detector of the array is provided with a first type of linear polarization filter and at least one second photo-sensitive detector is provided with a second type of linear polarization filter. The first type of linear polarization filter has a plane of polarization which is at angled with respect to a plane of polarization of said second type of polarization filter.

PRIORITY CLAIM

This application claims priority from European Application for PatentNo. 15201144.1 filed Dec. 18, 2015, the disclosure of which isincorporated by reference.

TECHNICAL FIELD

Some embodiments relate to an apparatus and in particular but notexclusively to an apparatus for use in luminescence applications, forexample, fluorescence imaging applications.

BACKGROUND

Various fluorescence applications are known such as fluorescenceanisotropy imaging microscopy (FAIM), and fluorescence lifetime imagingmicroscopy (FLIM).

Fluorescence anisotropy imaging microscopy (FAIM) concerns the study ofmolecular orientation and mobility using linearly polarized light. Thelinearly polarized light preferentially excites fluorophores which havea dipole orientation (absorption transition moment (ATM)) similar to theplane of the electric-field of the linearly polarized light. Thefluorophores which have a dipole orientation orthogonal to the plane ofthe electric-field are not excited, this is known as photo selection. Asthe fluorophores rotate due to their environment and undergo otherprocesses, the spontaneously emitted light becomes more depolarized. Thedegree of depolarization that has occurred can be measured by separatingthe emitted light into orthogonal linear components. The two maincontributing factors to the depolarization are rotational diffusion andFörster resonance energy transfer (FRET). This is a particularly usefultechnique as FAIM can therefore provide spatially resolved informationon rotational mobility, molecular binding, or clustering offluorescently labeled molecules, without dependence on signal intensity.

Anisotropy can be measured using both steady-state FAIM and/ortime-resolved FAIM.

With steady-state FAIM, the degree of anisotropy is measured by takingan average of the polarized fluorescence during the excitation time(i.e. exposure time). The steady state FAIM does not allow anyinformation into how the degree of anisotropy changes with respect totime during the excitation (i.e., exposure time). Steady state FAIM isuseful to compare cellular systems, as cells with a high degree ofproximity will present a lower average degree of anisotropy than cellswhich are further apart.

Time-resolved FAIM, allows the change in the degree of anisotropy to bemeasured with respect to time. Upon the excitation of a fluorophoretagged to a cell oriented in the same direction as the incidentpolarized light, the fluorophore will fluoresce, wherein thefluorescence has a high degree of polarization (i.e. indicating highanisotropy). If the same fluorophore starts coupling energy toneighboring fluorophores tagged to cells that are randomly oriented, thelatter fluorophores will start to fluoresce and emit light which has alower degree of polarization (i.e. indicating low anisotropy). As timepasses the degree of anisotropy will therefore decay.

Fluorescence lifetime imaging microscopy (FLIM) is an imaging techniquebased on differences in the average decay rate of excited states from afluorescent sample. The contrast in a FLIM image is thus based on thelifetime of individual fluorophores rather than their emission spectra.Unlike intensity measurements, fluorescence lifetime measurements do notdepend on: concentration, absorption by the sample, sample thickness,photo-bleaching, and/or excitation intensity.

The fluorescence lifetime of a fluorophore is the average decay rate ofexcited states from a fluorescent sample, and is characteristic for eachfluorescent molecule. As a result, it can be used to characterize asample. The fluorescent lifetime, however, is dependent on the localenvironment of the fluorophore, including: FRET, quenching, molecularrotation pH, ion or oxygen concentration, molecular binding or proximityof energy acceptors, as such it is possible to ascertain a wealth ofinformation from the fluorophore by measuring its lifetime. FLIM isoften used to observe a change, typically a reduction, in thefluorescence lifetime of a donor, when different fluorophores are inclose proximity.

FRET is a process which describes the non-radiative transfer of energybetween two similar energy systems that lie physically close together.For example, a donor fluorophore, which is initially in an excited statemay transfer energy to an acceptor fluorophore through non-radiativedipole-dipole coupling. In doing so, the acceptor enters an excitedstate, with the donor becoming quenched. The efficiency of this energytransfer is extremely sensitive to small changes between the distance ofthe donor and acceptor, and is inversely proportional to the sixth powerof that distance. This results in changes to the fluorescence intensityand the fluorescence lifetimes of the two fluorophores.

Two forms of FRET which are of importance are homo-FRET and hetero-FRET.In homo-FRET, only one type of fluorophore is present, as such theenergy transfer is reversible. This results in the fluorescent emissionform the fluorophore having largely the same polarization as that of theincident excitation beam. In hetero-FRET, two types of fluorophores arepresent (for example, A, B), as such the fluorophores become mixed intoa combination of pairs, for example AA, AB, BA, and BB. If theexcitation is tuned to the absorption peak of A, the fluorescenceconsists of contributions from A (homo-FRET), and B (hetero-FRET). Thehomo-FRET and hetero-FRET emission may be spectrally separated and thusthe hetero-FRET signal is more depolarized than the homo-FRET signal. Inhetero-FRET, the fluorescence lifetime of the donor changes as afunction of distance between the donor and acceptor, typically thecloser the acceptor is to the donor, the shorter the fluorescencelifetime of the donor.

Typically FAIM (both steady-state, and time-resolved) is used to measurehomo-FRET, while FLIM is typically used to measure hetero-FRET.

Currently, the systems used in this field are both bulky and expensive.Some embodiments aim to address or at least mitigate this.

SUMMARY

According to a first aspect there is provided a detector for detectingphotoluminescence from a sample, the detector comprising: an array ofphoto-sensitive detectors configured to receive photoluminescence, atleast one photo-sensitive detector being provided with a first type oflinear polarization filter and at least one photo-sensitive detectorbeing provided with a second type of linear polarization filter, whereinthe first type of linear polarization filter has a plane of polarizationwhich is angled with respect to a plane of polarization of the secondtype of polarization filter.

The first type of linear polarization filter may have a plane ofpolarization which is at or substantially at right angles to a plane ofpolarization of the second type of polarization filter.

The detector may be used to detect fluorescence.

Each photo-sensitive detector may be a SPAD.

Each photosensitive detector with one of said first and second linearpolarization filter is individually provided with said linearpolarization filter.

Each linear polarization filter may be provided by a grid of parallelwires, the longitudinal extent of said wires defining the plane ofpolarization.

The spacing between adjacent wires may be less than a wavelength oflight.

At least one photosensitive detector is provided with a wavelengthfilter.

A first plurality of photosensitive detectors may be provided with a redfilter, a second plurality of photosensitive detectors may be providedwith a green filter, and a third plurality of photosensitive detectorsmay be provided with a blue filter.

Each of said first, second, and third plurality of photosensitivedetectors comprise at least one photosensitive detector with the firsttype of linear polarization filter and at least one photosensitivedetector with the second type of linear polarization filter.

At least one photosensitive detector may be provided with no wavelengthfilter.

At least one photosensitive detector may be provided with no linearpolarization filter.

Each of said first, second, and third plurality of photosensitivedetectors comprise at least one photosensitive detector with no linearpolarization filter.

Each respective filter may cover more than one photosensitive detector.

The detector may comprise a signal processor configured to processoutputs from said array to provide, using the same detector,fluorescence anisotropy imaging microscopy FAIM information, andfluorescence lifetime imaging microscopy FLIM information.

The processor may be configured to correlate outputs from thephotosensitive detectors which have linear polarizers in FAIMtechniques, with outputs from the photosensitive detector which have nolinear polarizers.

The processor may be configured to correlate outputs from thephotosensitive detectors which have no linear polarizers in FLIMtechniques, with outputs from the photosensitive detectors which havelinear polarizers.

According to another aspect there is provided an integrated circuitcomprising the detector as discussed previously.

BRIEF DESCRIPTION OF THE DRAWINGS

Some embodiments will now be described by way of example only and withreference to the accompanying Figures in which:

FIG. 1A schematically shows the principles used in some embodiments;

FIG. 1B shows schematically the horizontal and vertical polarizationcomponents;

FIG. 2 schematically shows a system;

FIG. 3 schematically shows a second system;

FIG. 4 schematically shows a third system using a single detector;

FIG. 5A shows a cross section of a SPAD device used in some embodiments;

FIG. 5B shows a cross section of a second SPAD device used in someembodiments;

FIG. 6 shows a perspective view of the SPAD device of FIGS. 5A-5B;

FIG. 7 shows schematically an example of an imager with linearpolarization filtering;

FIG. 8 shows the imager of FIG. 7 with the addition of color filters andschematically shows the associated processing functions; and

FIG. 9 shows the method of ISP processing when the imager of FIG. 7 isused in the arrangement of FIG. 4.

DETAILED DESCRIPTION

Reference is made to FIG. 1A which shows the principles used in someembodiments. Light from a light source 100 is emitted and passes througha linear polarizer 101. The linear polarizer 101 will cause theunpolarized light emitted by the light source 100 to be linearlypolarized (Vertical-polarization in the example shown), V-polarization.The vertically polarized light 102 is then directed onto a sample 104,which for example, may be in a cuvette, or for example, may be on amicroscope slide. If, for example, the sample 104 is optically excitedat an appropriate wavelength (i.e., with a photon energy higher than thelowest unoccupied molecular orbital, or other similar mechanism) by thevertically polarized light 102, the sample 104 will subsequently emitlight 110 which is unpolarized. The light emitted from the sample 110,may for example be, fluorescence.

Reference is made to FIG. 1B, the optical emission (unpolarized light)110 emitted by sample 104, will have both vertical and horizontal planarcomponents. The intensity of the planar components (for examplevertical, or horizontal) of the light emitted 110 by the sample 104, mayprovide intracellular information of the sample 104.

Reference is now made to FIG. 2 which schematically shows a system.Light from the light source 100 is emitted and passes through a firstlinear polarizer 101. The linear polarizer 101 will cause theunpolarized light emitted by the light source 100 to be linearlypolarized, for example, vertically polarized. The vertically polarizedlight 102 is then directed via a first beam splitter 111 onto a sample104, for example through a microscope objective 103. The opticalemission 110 from the sample 104, which will be unpolarized, may then becollected by the microscope objective 103. The unpolarized opticalemission 110 may then be passed through the first beam-splitter 111. Thefirst beam-splitter 111 may for example have dichroic properties whichmay filter the wavelength of the optical emission 110, to a desiredwavelength range. In some embodiments a monochromator, or otherwavelength filtering apparatus may for example be used. The unpolarizedoptical emission 110 may then be passed through a second beam-splitter112, which may for example, have a 50:50 transmission to reflectionratio.

The reflected unpolarized optical emission 110 may then be passedthrough a linear polarizer 113. The linear polarizer 113, for example,may only transmit the vertically polarized components 114 of the opticalemission 110 from the sample 104. Accordingly, only the verticallypolarized components 114 of the optical emission 110 from the sample104, will reach the detector 115, and hence be measured.

The transmitted unpolarized optical emission 110 may then be passedthrough a linear polarizer 116. The linear polarizer 116, for example,may only transmit the horizontally polarized components 117 of theoptical emission 110 from the sample 104. Accordingly, only thehorizontally polarized components 117 of the optical emission 110 fromthe sample 104, will reach the detector 118, and hence be measured.

Reference is now made to FIG. 3 which shows a second system. In thisexample, the light 102 emitted from the light source 100 may be pulsed.If, for example, the sample 104 is excited with a pulsed light source100, the optical emission 110 from the sample 104 will also be pulsed.If the decay time of the pulsed light source 100 is sufficiently short(for example, less than a nanosecond), and the response time of thedetectors 115, and 118 is fast enough, the lifetime of the opticalemission 110 may be measured. This, for example, may be accomplishedusing a Time-Correlated Single Photon Counting (TCSPC) technique, usinga TCSPC unit 123. The laser driver 125 may trigger the light source 100to emit a pulse of light using a trigger signal 126. The laser drivermay in turn be controlled by the TCSP unit 123 using a control signal124. In some embodiments, other techniques may be used to pulse modulatethe light emitted from the source, for example an acoustic-opticalmodulator (AOM) in the beam path, or for example a Q-switched lasersystem, or similar. The pulse of light may then be directed onto thesample 104, with the respective vertical 114 and horizontal 117components, of the sample's 104 optical emission 110, being detected atdetectors 115 and 118 respectively as discussed above and reportingtheir respective detection data 121 and 122 to the TCSPC unit 123.Correlating the time at which the light source 100 was triggered by thelaser driver 125, with the optical emission 110 detected at detectors115 and 118 respectively, may allow the lifetime of the optical emission110 to be measured. It may also be possible, for example, to measure thelifetime of the vertical components 114, and the vertical components 117of the optical emission 110 separately.

For example, in using the techniques described above it may be possibleto measure the lifetime of both homo-Forster Resonance Energy Transfer(FRET) and hetero-Forster Resonance Energy Transfer (FRET) betweenspectrally different fluorophores. However, these techniques arecumbersome, and measuring the lifetime of the excited states of thefluorophores using FLIM, would require a third detector withoutpolarizers. More equipment would be needed to further separate thesesignals spectrally, such as a monochromator, or colored filters.

Reference is now made to FIG. 4, which shows a similar arrangement asdiscussed above but which uses, for example, only a single detector 28,which may be used for FAIM and FLIM. The signal from the detector, maybe passed to an image signal processor 74, which may process theinformation into a desirable output. If only a single detector 28 isused, the complexity of signal processing performed within the processor74 required to separate the signals is increased.

In some embodiments, the detector 28 may use single photon avalanchediodes (SPAD) to sense the reflected light. In general, an array ofSPADs are provided as a sensor in order to detect a reflected lightpulse or light. A photon may trigger an avalanche current in one or moreof the SPADs in an SPAD array. The avalanche current may signal anevent, namely that a photon of light has been detected.

SPADs operate as follows. At an initial time, the diode isreverse-biased to a voltage larger than its breakdown voltage. Thereception of a photon in the diode junction area starts an avalanchecurrent in the diode, which creates an electric pulse. The diode is thenbiased back to a voltage smaller than the breakdown voltage to quenchthe avalanche current, so that the SPAD may again react to the receptionof a photon. However, the diode must again be reverse-biased to avoltage larger than its breakdown voltage in order to react to anotherphoton. SPADs can currently be used in cycles having reactivationperiods shorter than 10 ns.

The detector 28 may comprise one or more SPADs. In some embodiments, anarray of SPADs will be provided. As will be discussed in more detaillater, some or all of the SPADs will have a linear polarizing filter 40.

Reference is made to FIG. 5A which schematically shows a cross sectionof a SPAD used in some embodiments. The SPAD 54 may be provided in ap-substrate 52. Respective p wells 30 and 42 may be provided in thesubstrate. Between the two p-wells 30 and 42 is provided an n-wellregion. The n-well region comprises a deep n-well 34, an n-well 32 andan n+ region 36. The n+ region 36 is adjacent the surface region of theSPAD and has a cathode 38 in contact therewith. Each of the p-wells 30and 42 is provided with a respective p+ region 46 and 44. A respectiveanode 48 and 50 is provided in contact with the respective p+ region 46and 44. It should be appreciated that a multiplication junction isprovided between the deep n-well 34 and the p-substrate 52.

Reference is now made to FIG. 5B which schematically shows a crosssection of a SPAD used in some embodiments. The SPAD 254 may be providedin a P-substrate 252. Respective P-wells 230, 231 and 232 may beprovided in the substrate. Each of the P-wells 230, 231 and 232 comprisea p+ region 234, 235 and 236. A respective anode 238 is provided to thep+ region 235. Each of the p+ regions 234 and 236 are provided withelectrical grounding 237 and 239. The N well regions 240 and 241 areprovided respectively between the P well regions 230 and 231 and betweenthe P well regions 23, and 232 (i.e. in a P-N-P-N-P arrangement). Eachof the N-wells 240 and 241 comprise an n+ region 242 and 243,respectively. The N wells 240 and 241 are provided with cathodes 244 and245, respectively. The N-P-N wells 240, 231 and 241, respectively, areprovided with a deep N-well 246. A multiplication junction 247 isprovided between the deep N-well 246 and P well 231. In some embodimentsthe linear polarizer grid edges 248 should contact from top metal downto Poly, to avoid side illumination.

A closely spaced metal wire grid 40 is provided over the surface of theSPAD and is thus between the SPAD detector and the source of thereflections. The wire grid has a plurality of parallel wires. Theclosely spaced wire grid acts as a polarization filter for light. Thespacing d between the wires may be smaller than the wavelength of lightand may for example be: d=λ/2 (where λ is wavelength). The width w ofthe wire may be as small as the process allows: w=minimum (where w isthe wire width, where the wire may for example be made of metal), thevalue of w may for example be 0.14 μm.

In some embodiments, a wire grid filter may be provided on eachindividual SPAD.

It should be appreciated that the orientation of the wires controls thedirection of the linear polarization provided by the grid.

In this regard, reference is made to FIG. 6 which schematically showsthe grid 40 provided on the SPAD 54. In some embodiments, it is ensuredthat the grid end edges, in the longitudinal direction parallel to thewires, should contact from the top metal down to the poly layer to avoidside illumination impinging on the SPAD, thus bypassing the polarizationfunction provided by the grid. The edges which are perpendicular to thelongitudinal direction of the wires do still provide a polarizationfunction. However, in some embodiments, the grid end edges in theparallel direction to the wires may be such that there is a continuousside edge to prevent any side illumination.

Reference is now made to FIG. 7 which schematically shows an arrangementof filters on the SPAD array. In this example, some of the SPADs 60 arewithout any wire grid and are thus able to detect all light includingpolarized light. In other words, these SPADs do not have any filter onthem. Some of the SPADs 62 are each provided with a wire grid which isoriented in one direction and some of the SPADs 64 are provided withwire grids which are oriented in a direction perpendicular to that ofSPADs 62. In the example shown in FIG. 7 those SPADs referenced 62 mayfor example only detect vertically polarized light whereas as thosewhich are referenced 64 are able to detect only horizontally polarizedlight.

Reference is made to FIG. 8 which shows a modification to thearrangement shown in FIG. 7. In this example, color filters areadditionally provided to allow spectral discrimination. For example,those pixels referenced 66 may have no color filter. The pixels withouta color filter 66 may be used to detect spectrally wide band emission(i.e., detect all of the emitted light without spectral filtering). Insome embodiments, these pixels 66 may be in an array together with RGBfiltered SPADs. In other embodiments, these pixels 66 may be usedexclusively.

Those pixels referenced 68 may have a green filter 69, those pixelsreferenced 70 may have a red filter 71 and those pixels referenced 72may have a blue filter 73. It should be appreciated that in differentembodiments more or less than the three color filters may be provided.It should be appreciated that alternatively or additionally, differentcolored filters may be used. In some embodiments, the color filters maybe provided on an individual pixel basis. Different patterns andpositions for the filters may thus be provided in different embodiments.

In some embodiments, the SPADs which have a linear polarizer in-front ofthem are used to measure homo-FRET in both steady-state andtime-resolved FAIM techniques. In other embodiments, the SPADs which donot have a linear polarizer in-front of them may be used in combinationor otherwise with the detected polarized light in FAIM techniques, forexample as photon counters.

In some embodiments, the SPADs which do not have a linear polarizerin-front of them are used to measure the fluorescence lifetime(hetero-FRET) in FLIM techniques. In other embodiments, the SPADs whichdo have a linear polarizer in-front of them may be used in combinationor otherwise with the detected unpolarized light in FLIM techniques, forexample as photon counters.

In some embodiments, SPADs with color filters in-front of them are usedto obtain spectrally distinct fluorescence of both homo-FRET, andhetero-FRET, using FAIM and FLIM techniques respectively. In someembodiments these color filters may be red, green, and blue. In otherembodiments other types of wavelength filters may be used, for example,dichroic filters, band pass filters, edge filters, notch filters, or thelike. In other embodiments SPADs with no color filters in-front of themare used to obtain spectrally wide band emission for both FAIM and FLIMtechniques.

Table 1 below shows an example of some embodiments, where an ‘X’indicates how information from the detector may be used. ‘Pol’ indicatesthat polarization information has been measured, and ‘none’ indicatesthat no polarization information has been measured.

TABLE 1 Red filter Green filter Blue filter No-color filter Pol none Polnone Pol none Pol none FAIM X X X X (Homo- FRET) FLIM X X X X (Hetero-FRET)

In some embodiments, information from the SPADs which measure polarizedemission are used to measure FAIM, and information from the SPADs whichdo not measure polarized emission are used to measure FLIM.

It should be appreciated that the information from at least onedetector, with or without a filter arrangement, may be used tocontribute information to the FAIM and FLIM techniques, using datacorrelation techniques.

The outputs from the SPAD array is provided to an image signal processor(ISP) 74. The image signal processor is thus able to provide outputs forthe vertically polarized light, the perpendicularly polarized light andthe fluorescence lifetime. In the ISP 74 the fluorescence lifetime iscalculated by fitting the time-resolved fluorescence decay, using FLIM,to an exponential function, wherein the lifetime is equal to theexponential decay constant of the exponential fit. This information isprovided for each of the three colors, as well as the wide band pixels.As can be seen, information for each of the different colors may begraphically represented for each of the polarizations. For example,graphs may be provided which show intensity plotted against wavelength(λ). The peaks referenced 80 are for the blue filtered results, thosereferenced 82 are for the green filtered results and those referenced 84are for the red filtered results. The first graph shows the results forthe vertical polarization, the second graph shows the results for thehorizontal polarization and the third graph shows the results wherethere is no polarization filter.

Reference is now made to FIG. 9, which shows, for example, theprocessing undertaken by the image signal processor (ISP) 74. The signalgenerated by the detector depicted in FIG. 8 by references 66, 68, 70and 72, may then be passed into the ISP 74 as an input from the detectorarray 91. The timing signal from the laser driver may then also bepassed into the ISP 74 as an input from the laser driver 92. The ISP 74,may then separate 93 the spectral, and polarized and un-polarizedcomponents of the signal 91 from the detector. This information may thenbe temporally correlated 94 with the input from the laser driver 92. TheISP 74 may then output the polarized information and lifetime components76, 77, and 78. The polarized information and lifetime components maythen be further separated into their spectral components for example,using the red, green and blue color filters described earlier. Forexample, the parallel polarized lifetime output 76, may be separatedinto red 76 a, green 76 b, and blue 76 c. The orthogonal polarizedlifetime output may be separated into red 77 a, green 77 b, and blue 77c. Additionally, the unpolarized lifetime may be separated into red 78a, green 78 b, and blue 78 c.

It should be appreciated that the information which is captured by thearray can be processed in any suitable manner.

It should be appreciated that some embodiments of the invention mayallow a compact device to the achieved.

In some embodiments the polarization filters, and/or color filters, maybe printed onto one or more transmissive surfaces, for example, silicaor quartz, to form an arrayed optical filter. Such an optical filter maythen be positioned in-front of an array of SPADs, to achieve a similareffect.

In some embodiments the relationship between filters and photo-sensitivedetectors is not one to one. A single filter, for example, a linearpolarizer, may cover more than one SPAD, for example four SPADs.Likewise, for example, a single color filter may cover more than oneSPAD. It should be appreciated that the number of SPADs, covered or notcovered, by at least one of the filters, may vary.

Some embodiments may use other photo-sensitive detectors, instead ofSPADs, for example, APDs, photodiodes, or the like. These sensors may beintegrating elements generating events on reception of the lightinformation.

It should be appreciated that the above described arrangements may alsobe used for other photoluminescence applications, for examplephosphorescence applications, or Raman spectroscopy. Raman spectroscopytypically being a method of characterizing a substance by reflectinglight of a known wavelength from a surface, and measuring small changesin the wavelength of the reflected light.

It should be appreciated that the above described arrangements may beimplemented at least partially by an integrated circuit, a chip set, oneor more dies packaged together or in different packages, discretecircuitry or any combination of these options.

Various embodiments with different variations have been described hereabove. It should be noted that those skilled in the art may combinevarious elements of these various embodiments and variations.

Such alterations, modifications, and improvements are intended to bepart of this disclosure, and are intended to be within the scope of thepresent invention. Accordingly, the foregoing description is by way ofexample only and is not intended to be limiting. The present inventionis limited only as defined in the following claims and the equivalentsthereto.

The invention claimed is:
 1. A detector for detecting photoluminescencefrom a sample, said detector comprising: an array of photo-sensitivedetectors configured to receive photoluminescence, at least one firstphoto-sensitive detector in said array having a first type of linearpolarization filter and at least one second photo-sensitive detector insaid array having a second type of linear polarization filter, whereinsaid first type of linear polarization filter has a plane ofpolarization which is at angled with respect to a plane of polarizationof said second type of polarization filter; each photo-sensitivedetector having one of said first and second linear polarization filtersis individually provided with a linear polarization filter.
 2. Thedetector as claimed in claim 1, wherein the detector is used to detectfluorescence.
 3. The detector as claimed in claim 1, wherein eachphoto-sensitive detector is a SPAD.
 4. The detector as claimed in claim1, wherein each of the first and second linear polarization filters isprovided by a grid of parallel wires, wherein a longitudinal extent ofsaid parallel wires defining a plane of polarization.
 5. The detector asclaimed in claim 4, wherein a spacing between adjacent wires in the gridof parallel wires is less than a wavelength of light to be detected. 6.The detector as claimed in claim 1, wherein at least one photo-sensitivedetector of said array has a wavelength filter.
 7. The detector asclaimed in claim 6, wherein first ones of the plurality ofphoto-sensitive detectors in said array are provided with a redwavelength filter, second ones of the plurality of photo-sensitivedetectors in said array are provided with a green wavelength filter, andthird ones of the plurality of photo-sensitive detectors in said arrayare provided with a blue wavelength filter.
 8. The detector as claimedin claim 7, wherein each of said first, second, and third ones of theplurality of photo-sensitive detectors comprise said firstphoto-sensitive detector having the first type of linear polarizationfilter and said second photo-sensitive detector having the second typeof linear polarization filter.
 9. The detector as claimed in claim 8,wherein at least one third photo-sensitive detector of said array isprovided with no linear polarization filter.
 10. The detector as claimedin claim 9 wherein each of said first, second, and third ones of theplurality of photo-sensitive detectors further comprise said thirdphoto-sensitive detector of said array with no linear polarizationfilter.
 11. The detector as claimed in claim 7, wherein fourth ones ofthe plurality of photo-sensitive detectors in said array do not have awavelength filter.
 12. A detector for detecting photoluminescence from asample, comprising: an array of photo-sensitive detectors configured toreceive photoluminescence, at least one first photo-sensitive detectorin said array having a first type of linear polarization filter and atleast one second photo-sensitive detector in said array having a secondtype of linear polarization filter, wherein said first type of linearpolarization filter has a plane of polarization which is at angled withrespect to a plane of polarization of said second type of polarizationfilter; and a signal processor configured to process outputs from saidarray to provide both fluorescence anisotropy imaging microscopy (FAIM)information and fluorescence lifetime imaging microscopy (FLIM)information.
 13. The detector as claimed in claim 12, wherein saidsignal processor is configured to correlate outputs from thephoto-sensitive detector which have linear polarizers in FAIM techniqueswith outputs from the photo-sensitive detector which have no linearpolarizers.
 14. The detector as claimed in claim 12, wherein said signalprocessor is configured to correlate outputs from the photo-sensitivedetector which have no linear polarizers in FLIM techniques with outputsfrom the photo-sensitive detectors which have linear polarizers.
 15. Adetector for detecting photoluminescence from a sample, said detectorcomprising: an array of photo-sensitive detectors configured to receivephotoluminescence, said array including a first set of photo-sensitivedetectors and a second set of photo-sensitive detectors; a coloredfilter over the second set of photo-sensitive detectors, wherein thereis no colored filter over the first set of photo-sensitive detectors;wherein each of first and second sets of photo-sensitive detectorsincludes: a first photo-sensitive detector having a first type of linearpolarization filter; and a second photo-sensitive detector having asecond type of linear polarization filter; wherein the first and secondtypes of linear polarization filters have different planes ofpolarization.
 16. The detector of claim 15, wherein each of first andsecond sets of photo-sensitive detectors further includes a thirdphoto-sensitive detector having no polarization filter.
 17. The detectorof claim 16, wherein the colored filter is selected from the groupconsisting of a red wavelength filter, a blue wavelength filter and agreen wavelength filter.
 18. The detector of claim 16, furthercomprising a signal processor configured to process outputs from saidarray of photo-sensitive detectors to provide both fluorescenceanisotropy imaging microscopy (FAIM) information and fluorescencelifetime imaging microscopy (FLIM) information.
 19. The detector ofclaim 18, wherein said signal processor is configured to correlateoutputs from the first and second photo-sensitive detectors of saidarray which have linear polarization filters in FAIM techniques withoutputs from the third photo-sensitive detectors of said array whichhave no polarization filter.
 20. The detector of claim 18, wherein saidsignal processor is configured to correlate outputs from the thirdphoto-sensitive detectors of said array which have no polarizationfilter in FLIM techniques with outputs from the first and secondphoto-sensitive detectors of said array which have linear polarizationfilters.
 21. A detector for detecting photoluminescence from a sample,said detector comprising: an array of photo-sensitive detectorsconfigured to receive photoluminescence, at least one firstphoto-sensitive detector in said array having a first type of linearpolarization filter and at least one second photo-sensitive detector insaid array having a second type of linear polarization filter, whereinsaid first type of linear polarization filter has a plane ofpolarization which is at angled with respect to a plane of polarizationof said second type of polarization filter; and wherein each of thefirst and second linear polarization filters is provided by a grid ofparallel wires, a longitudinal extent of said parallel wires defining aplane of polarization.
 22. The detector of claim 21, wherein a spacingbetween adjacent wires in the grid of parallel wires is less than awavelength of light to be detected.
 23. The detector of claim 21,wherein the detector is used to detect fluorescence.
 24. The detector ofclaim 21, wherein each photo-sensitive detector is a SPAD.
 25. Thedetector of claim 21, wherein at least one photo-sensitive detector ofsaid array has a wavelength filter.
 26. The detector of claim 25,wherein first ones of the plurality of photo-sensitive detectors in saidarray are provided with a red wavelength filter, second ones of theplurality of photo-sensitive detectors in said array are provided with agreen wavelength filter, and third ones of the plurality ofphoto-sensitive detectors in said array are provided with a bluewavelength filter.
 27. The detector of claim 26, wherein each of saidfirst, second, and third ones of the plurality of photo-sensitivedetectors comprise said first photo-sensitive detector having the firsttype of linear polarization filter and said second photo-sensitivedetector having the second type of linear polarization filter.
 28. Thedetector of claim 27, wherein at least one third photo-sensitivedetector of said array is provided with no linear polarization filter.29. The detector of claim 28, wherein each of said first, second, andthird ones of the plurality of photo-sensitive detectors furthercomprise said third photo-sensitive detector of said array with nolinear polarization filter.
 30. The detector of claim 26, wherein fourthones of the plurality of photo-sensitive detectors in said array do nothave a wavelength filter.